JP4116057B2 - Biomechanical stimulus loading device - Google Patents

Description

The present invention relates to a structure containing cultured cells, cultured tissues, or cultured cells in vitro culture using stimuli similar to vertical load stress stimuli and lateral shear stress stimuli generated in tissues in vivo on the ground ( In the present specification, the present invention generally relates to a biomechanical stimulus loading device for applying to a culture as a biomechanical stimulus repeatedly.

In recent years, as a treatment for patients with cartilage lesions, the remaining healthy cartilage tissue of the patient himself is collected, taken out of the body, separated from chondrocytes, and proliferated in in vitro culture. Cell transplantation treatment methods for transplantation to patients have been performed in Europe and the United States.
However, this method involves culturing and proliferating chondrocytes outside the body, but does not regenerate cartilage tissue outside the body, and its therapeutic effect is unstable, and it takes a long time to repair and mature the cartilage tissue. There was a problem that it took.

  In vivo tissues and cells are not only biochemically stimulated but also mechanically stimulated by gravity, muscle contraction force, and external force. Cultured cells and tissues are static or dynamic biomechanical stimuli. It is known that functions such as proliferation, differentiation, and metabolism are changed by receiving the drug.

  By the way, based on such knowledge, recently, cell culture apparatuses that perform cell culture and tissue culture using biomechanical stimulation have been studied and implemented by several research groups in and outside Japan (for example, Patent Document 1). reference).

  As a culture system using stress stimulation, a method of culturing chondrocytes or vascular smooth muscle cells by expanding or contracting or vibrating a membrane member to which cells are attached, or a chondrocyte held on a support in a column and pumping There is a method of culturing chondrocytes while circulating a medium and stimulating stress by flow of a culture solution to cartilage in a support.

  In addition, as a culture system using hydrostatic pressure stimulation, there is a gas cylinder type pressure loading method of chondrocytes, and a method of applying a pressure of 5 MPa corresponding to the internal pressure of the column to the column with a hydraulic cylinder pump is applied to the chondrocytes. I'm trying to do a stimulus load.

  In addition, a cell culture device has been proposed in which a medium is pumped into a column (in-column device) and a pressure load pattern or cycle is arbitrarily created by changing the flow rate of the medium by operating a valve or the like. Yes.

  These methods and devices reproduce the pressure level applied to the cells in the living body, the pressure change, the pattern of pressure rise and fall, etc., close to the living body through the culture solution, The growth of cultured cells, the flow of the culture solution, and the stimulation of the cultured cells via the culture solution, and it cannot be said that the vertical biomechanical stimulation due to gravity on the supporting tissue such as bone and cartilage has been reproduced. It was a thing.

Here, vertical vertical stimulation by gravity is important for the living body when healthy astronauts live in a microgravity space and the stimulation by gravity decreases or they are forced to lie down due to illness or age. It is also clear from osteoporosis in the spine and upper and lower limbs.
JP 2003-265164 A

As described above, the above-described conventional technique does not reproduce the load applied in the vertical direction due to the gravity of the cultured cells and the force applied to the cells in the state surrounded by the matrix that is the extracellular tissue.
The present invention provides a structure containing cultured cells, cultured tissues, or cultured cells in vitro culture by applying a stimulus similar to a vertical load stress stimulus or a lateral shear stress stimulus generated in the tissue in vivo on the ground. An object of the present invention is to provide a biomechanical stimulus loading device for applying repeatedly as a biomechanical stimulus.

In order to achieve the above object, the biomechanical stimulus load device of the present invention is a biomechanical stimulus load device comprising a mechanical stimulus load device and a culture vessel installed in a carbon dioxide incubator, and a control computer. The mechanical stimulation load device includes a load load piston, a piston vertical movement stage that supports the load load piston to be movable in a vertical direction within a predetermined range, and the piston vertical movement stage in the vertical direction. A stage elevating mechanism for moving the piston up and down, and by driving the stage elevating mechanism in a predetermined cycle by a control computer, the piston vertical movement stage is moved up and down, and when the piston vertical movement stage descends, biomechanical stimulation load devices weight of the load the load piston is rests on the culture in the culture vessel The piston mounting stage is mounted on the piston vertical movement stage, and the load loading pistons are respectively inserted into and supported by the plurality of holes formed in the piston mounting stage. And a common hole for allowing the plurality of load-loading pistons to freely move in the vertical direction is formed in the piston vertical movement stage .

  In this case, an additional load weight can be attached to the upper end portion of the load load piston.

  Further, a pressurizing body having a shape corresponding to the culture body can be attached to the lower end portion of the load loading piston.

  Further, the piston mounting stage configured to support a plurality of load-loading pistons can be configured to be movable in a horizontal plane with respect to the piston vertical movement stage.

  Further, when the load loading piston moves in the vertical direction, it can be supported by the piston vertical movement stage or the piston mounting stage so as not to rotate.

  Further, the culture container can be detachably attached to the culture container fixing stage of the mechanical stimulation load device.

  In addition, a shear stress loading mechanism for moving or vibrating the culture vessel in a horizontal plane can be provided.

  In addition, it is possible to fix the receiving group having a predetermined shape in the culture container, and the culture body load-load stimulating portion including the pressurizing body and the receiving group at the lower end of the load-loading piston has a predetermined load such as a knee joint. A structure similar to the shape can be obtained.

  In addition, a member made of a biocompatible synthetic resin material can be attached to the culture body load-stimulating portion composed of the pressurizing body and the receiving group at the lower end of the load-loading piston.

According to the cell culture method using a biomechanical stimulus load by the biomechanical stimulus load device of the present invention, the weight of the load-loading piston is applied to the culture body in the culture vessel in a predetermined cycle, whereby the culture in the culture vessel is performed. Since a vertical load load stimulus is applied to the body, a stimulus similar to the vertical load load stimulus generated in the tissue in vivo on the ground can be applied to the culture body. Since it can be cultured in a state that approximates the environment of the tissue in the living body, it becomes possible to create a differentiated and mature culture that can withstand the mechanical load after transplantation, and to efficiently repair and mature the tissue of the living body Can do.

  In addition, a shear stress stimulus in the lateral direction is applied by moving or vibrating the culture container in which the culture body is placed in a horizontal plane while applying a vertical load load stimulus to the culture body in the culture container. By doing so, the culture body can be stimulated in the same manner as the lateral shear stress stimulation generated in the tissue in vivo on the ground, and the culture body is cultured in a state approximating the tissue environment in the body. Therefore, it is possible to create a differentiated and matured culture that can withstand the mechanical load after transplantation, and more efficiently repair and mature the tissue of the living body.

Then, according to the biomechanical stimulation load device of the present invention, mechanical irritation load device, and a piston for load application, the piston vertically mobile for movably supporting該荷heavy load piston in the vertical direction within a predetermined range A stage and a stage lifting mechanism for moving the piston vertical movement stage in the vertical direction, and the piston vertical movement stage is moved in the vertical direction by driving the stage lifting mechanism in a predetermined cycle by a control computer. Since the weight of the load loading piston is applied to the culture body in the culture vessel when the piston vertical movement stage is lowered, the vertical direction generated in the tissue in the living body on the ground An apparatus for accurately applying a stimulus similar to a load load stimulus to a culture can be obtained easily and at low cost.
Here, the piston mounting stage is mounted on the piston vertical movement stage, and the load loading pistons are respectively inserted into and supported by the plurality of holes formed in the piston mounting stage. A plurality of load-loading pistons can be freely moved in the vertical direction on the piston vertical movement stage, thereby forming a vertical load load stimulus on the plurality of culture bodies. Can be applied simultaneously with the same or different magnitude of the load load stimulus, the culture can be efficiently cultured, and the support of various load load pistons is simplified. Can be done.

  In addition, by attaching an additional load weight to the upper end of the load load piston, the magnitude of the vertical load load stimulus applied to the culture can be adjusted accurately and easily.

  In addition, by attaching a pressurizing body having a shape corresponding to the culture body to the lower end of the load-loading piston, the same stimulation as the vertical load-loading stimulation generated in the tissue in vivo on the ground It can be accurately applied to the culture.

  In addition, by arranging a piston mounting stage that supports a plurality of load-loading pistons so that it can move in a horizontal plane with respect to the piston vertical movement stage, Different vertical loads can be applied.

  Further, when the load-loading piston moves in the vertical direction, it is supported on the piston vertical movement stage or the piston mounting stage so that it does not rotate, so that the culture body is placed on the lower end of the load-loading piston. Even when a pressurizing body having a shape corresponding to the above is mounted, a stimulus similar to a vertical load load stimulus generated in a tissue in a living body on the ground can be accurately applied to the culture body.

  Moreover, by detachably mounting the culture container on the culture container fixing stage of the mechanical stimulation load device, various culture containers can be easily mounted.

  In addition, by providing a shear stress loading mechanism that moves or vibrates the culture vessel in a horizontal plane, a device that accurately applies to the culture body the same stimulus as the lateral shear stress stimulus generated in the tissue in vivo on the ground Can be obtained easily and at low cost.

  In addition, it is possible to fix the receiving group having a predetermined shape in the culture container, and the culture body load-load stimulating portion including the pressurizing body and the receiving group at the lower end of the load-loading piston has a predetermined load such as a knee joint. By making the structure similar to the shape, the force applied to the cells in the state surrounded by the matrix that is the extracellular tissue can be easily reproduced.

  In addition, by attaching a member made of a biocompatible synthetic resin material to a culture body load-stimulating portion composed of a pressurizing body and a receiving group at the lower end of the load-loading piston, Can be cultured in a state approximating the environment of the tissue in the living body, so that the adaptability of the culture body to the living body can be improved.

Embodiments of the biomechanical stimulation load device of the present invention will be described below with reference to the drawings.

In FIGS. 1-2, showing a first embodiment of a biomechanical stimulation load device of the present invention.

This biomechanical stimulation load device is composed of a mechanical stimulation load device 2 and a culture vessel 3 installed in a carbon dioxide incubator 1 and a control computer 4.
In addition, the biomechanical stimulation load device can be operated from outside the carbon dioxide incubator 2, thereby enabling culturing for a long time while maintaining the sterilized state in the carbon dioxide incubator 2. Can do.

  The inside of the carbon dioxide gas incubator 2 is sterilized and used in an environment in which temperature, humidity, oxygen, carbon dioxide, nitrogen partial pressure and the like are controlled.

The mechanical stimulation load device 2 installed in the carbon dioxide incubator 1 includes a load-loading piston 5 and a piston vertical movement stage 21 that supports the load-loading piston 5 so as to be movable in a vertical direction within a predetermined range. And a stage elevating mechanism 22 for moving the piston up / down moving stage 21 in the up / down direction, and the stage elevating mechanism 22 is driven in a predetermined cycle by the control computer 4 to thereby move the piston up / down moving stage 21. Are moved in the vertical direction, and the weight of the load-loading piston 5 is applied to the culture body C in the culture vessel 3 when the piston vertical movement stage 21 is lowered.
Thereby, it is possible to give the culture body C the same stimulus as the vertical load load stimulus generated in the tissue in the living body on the ground.

The stage lifting mechanism 22 for moving the piston vertical movement stage 21 in the vertical direction includes, for example, an electric actuator 22a and a wire 22b that is connected to the electric actuator 22a and suspends the piston vertical movement stage 21.
Then, by operating the electric actuator 22a in accordance with a command from the control computer 4, the piston vertical movement stage 21 can be moved in the vertical direction along the guide member 20 via the wire 22b.

Incidentally, in this embodiment, through the piston mounting stage 23 placed on the piston vertically moving stage 21, a plurality number (e.g., five total of 25 five ×) load load piston 5 I try to support it.
In this case, since the piston vertical movement stage 21 is for mounting the piston mounting stage 23, a hole for allowing the load-loading piston 5 to freely move in the vertical direction at the center. The part 21a is formed.
Thereby, various pistons 5 for load loading can be supported simply.
As a reference example, the piston mounting stage 23 may be omitted, and the load loading piston 5 may be directly supported on the piston vertical movement stage 21.

The load-loading piston 5 has a pressurizing portion 51 having a shape corresponding to the culture C at the lower end portion, and a guide shaft portion 52 fitted in a hole portion 23a formed in the piston mounting stage 23 in the intermediate portion. A large diameter portion 53 is formed at the upper portion, and a heavy load attaching portion 54 for attaching an additional load heavy load 55 is formed at the upper end portion.
As a result, as shown in FIG. 2A, the piston vertical movement stage 21 is moved downward from the state in which the load loading piston 5 is supported on the piston mounting stage 23, and the state shown in FIG. As shown, when the piston vertical movement stage 21 is lowered, the load loading piston 5 is released from the support on the piston mounting stage 23 by the large-diameter portion 53 of the load loading piston 5 so that the weight of the load loading piston 5 is increased. It is applied directly to the culture body C in the culture vessel 3.
Thereafter, the piston vertical movement stage 21 is moved upward to return the piston loading stage 23 to the state where the piston 5 for load loading is supported, as shown in FIG.
The load load stimulus in the vertical direction by the load load piston 5 can freely operate the load cycle and the load time by the operation of the control computer 4.
In addition, the additional load heavy load 55 can be attached to the heavy load attachment portion 54 as necessary, and the weight thereof can be freely set, so that the magnitude of the vertical load load stimulus applied to the culture C is large. Can be easily adjusted.

Moreover, the shape of the pressurizing portion 51 of the load-loading piston 5 can be any shape corresponding to the culture body C, and further, as in the modification of the load-loading piston 5 shown in FIG. The pressurizing body 51A having a shape corresponding to the body C may be formed as a separate member and attached to the lower end 51B of the load-loading piston 5.
Thereby, the stimulus similar to the vertical load load stimulus generated in the tissue in the living body on the ground can be accurately given to the culture body C.

Further, the load loading piston 5 can be supported by the piston placement stage 23 (or the piston vertical movement stage 21) so as not to rotate when moving in the vertical direction.
Specifically, the hole portion 23a formed in the piston mounting stage 23 is formed in a polygonal shape, and the guide shaft portion 52 is formed so as to have a polygonal cross-sectional shape adapted to the polygonal hole portion 23a. .
Thereby, even when the pressurizing body 51A having a shape (other than a circle) corresponding to the culture body C is attached to the lower end portion 51B of the load-loading piston 5, the vertical generated in the tissue in the living body on the ground. The stimulus similar to the direction load load stimulus can be accurately applied to the culture C.

The culture vessel 3 is detachably attached to the culture vessel fixing stage 24 of the mechanical stimulus loading device 2.
Thereby, various culture vessels 3 can be easily attached.
In addition, a culture vessel (for injection) 32 and a culture vessel (for discharge) 33 are connected to the culture container 3 as necessary, and the injection and discharge of the culture medium are controlled by the control computer 4. You can also

In addition, a shear stress loading mechanism 26 that moves or vibrates the culture vessel 3 in a horizontal plane is provided.
Specifically, a shear stress stage 25 is disposed on the culture vessel fixing stage 24 so as to be movable in a horizontal plane with respect to the culture vessel fixing stage 24, and the culture vessel 3 is mounted on the shear stress stage 25. In addition, the shear stress stage 25 is moved or vibrated in a horizontal plane by the shear stress loading mechanism 26.
Note that the direction of movement or vibration in the horizontal plane is not limited to one direction, and can be any direction such as two directions in the X and Y directions and circular motion.
As the shear stress load mechanism 26, for example, an electric actuator or a moving mechanism or a vibration mechanism combining a permanent magnet and an electromagnet can be used. The drive of the shear stress load mechanism 26 is controlled by the control computer 4. To do.
Thereby, a lateral shear stress stimulus generated in the tissue in the living body on the ground, for example, a stimulus similar to a stimulus applied at the time of bending and extending motions to the knee joint can be applied to the culture body C.
In this case, the culture vessel 3 is moved or vibrated in the horizontal plane by the shear stress loading mechanism 26 while adjusting the method of applying the vertical load load stimulus to the culture body C in the culture vessel 3 by the load loading piston 5. Accordingly, it is possible to apply shear stress stimulation in an oblique direction.

As shown in FIGS. 2B and 4, an internal culture container 30 having an arbitrary shape suitable for the culture body C is installed in the culture container 3, and the load-loading piston 5 of the internal culture container 30 is installed. A receiving group 31 of a structure including a cultured cell, a cultured tissue or a cultured cell is mounted at a position facing the pressurizing unit 51, and the cultured cell, the cultured tissue or the cultured cell as the cultured body C is included on the receiving group 31. A structure is placed.
The pressurizing body 51A of the load-loading piston 5 shown in FIG. 3 and the flat-shaped receiving group 31 of the internal culture container 30 shown in FIG. 4 are suitable for the culture tissue of the skin as the culture body C. A vertical load stress stimulus and a transverse shear stress stimulus can be applied to the two, respectively.

In FIGS. 5-6, showing a second embodiment of the biomechanical stimuli loading device of the present invention.

  This biomechanical stimulation load device supports a plurality (plural types) of load-loading pistons 5a and 5b (pressurizing bodies 51Aa and 51Ab) on the biomechanical stimulation load device of the first embodiment. The piston mounting stage 23 is configured to be movable in a horizontal plane with respect to the piston vertical movement stage 21, and further, a plurality (a plurality of types) of additional load weights 55a and 55b are supported. The additional load heavy load mounting stage 28 is configured to be movable in a horizontal plane with respect to the additional load heavy load vertical movement stage 27, and the load load pistons 5a and 5b and the additional load heavy load loads 55a and 55b. Is added to the function that allows the user to select any combination.

Specifically, a piston mounting stage 23 is disposed on the piston vertical movement stage 21 so as to be movable (for example, rotatable) in a horizontal plane with respect to the piston vertical movement stage 21, and similarly added. An additional load weight reduction stage 28 is disposed on the load weight reduction vertical movement stage 27 so as to be movable (for example, rotatable) in a horizontal plane with respect to the additional load weight reduction vertical movement stage 27, for piston placement. The stage 23 and the additional load heavy restraint mounting stage 28 are independently moved (for example, rotated) in the horizontal plane by the piston mounting stage drive mechanism 23b and the additional load heavy restraint mounting stage drive mechanism 28b. To do.
As the piston mounting stage drive mechanism 23b and the additional load heavy load mounting stage drive mechanism 28b, for example, a drive mechanism combining an electric motor and a gear can be used. The driving of the load heavy restraint stage driving mechanism 28b is controlled by the control computer 4.

As shown in FIGS. 5 and 6, the load loading pistons 5a and 5b are supported on the piston placement stage 23, and the additional load weight restraints 55a and 55b are supported on the additional load weight placement stage 28, respectively. From the state, the piston vertical movement stage 21 and the additional load heavy restraint placement stage 28 are moved downward, and when the piston vertical movement stage 21 and the additional load heavy restraint placement stage 28 are lowered, the load load piston 5 The support for the piston mounting stage 23 by the large-diameter portion 53 and the support for the additional load heavy restraint 55a, 55b to the additional load heavy restraint mounting stage 28 are sequentially released so that the load load piston is released. The weights 5a and 5b and the additional load weights 55a and 55b are directly applied to the culture body C in the culture vessel 3.
Thereafter, by moving the piston up / down movement stage 21 and the additional load weight reduction stage 28 upward, as shown in FIGS. 2 and 6, the piston placement stage 23 is loaded with the load loading pistons 5a, 5b. In addition, the additional load heavy weights 55a and 55b are returned to the supported state on the additional load heavy weight loading stage 28, respectively.
Accordingly, a combination of the load-loading pistons 5a and 5b and the additional load weights 55a and 55b is arbitrarily selected, and different load-loading pistons 5a and 5b (pressurizing bodies 51Aa and 51Ab) are added to one culture body C. Thus, it is possible to easily apply different load loads in the vertical direction.

  The other configuration and operation of this biomechanical stimulus load device are the same as those of the biomechanical stimulus load device of the first embodiment.

7 shows a third embodiment of the biomechanical stimuli loading device of the present invention.

This biomechanical stimulation load device is suitably used for culturing meniscus, articular lips, joint discs, etc., and an internal culture container 30 suitable for the culture C is installed in the culture container 3, and the internal A receiving group 31 of a structure containing a cultured cell, a cultured tissue or a cultured cell is attached to a position opposite to the pressurizing body 51C of the load-loading piston 5 of the culture vessel 30. A structure including a cultured cell, a cultured tissue or a cultured cell is placed.
The meniscus, joint lip, joint disc, etc. are not stimulated in vivo with a load similar to that of bone, but are subjected to a load stimulus in osteochondral so that their culture (here, meniscus) ), An artificial femur bone cartilage 51D made of a synthetic resin of a biocompatible material similar to the femur bone cartilage is attached to the pressurizing body 51C of the load loading piston 5, and the tibial bone cartilage is attached to the receiving group 31. By placing the artificial tibial cartilage 31A made of a synthetic resin of a biocompatible material similar to the above, the culture body load load stimulating portion X formed therebetween becomes a predetermined shape of the living body (here, knee joint) The culture body C (here, a special meniscus culture dish for meniscus) is placed in the culture body load load stimulating portion X, and bone cartilage (here, osteoarticular cartilage of the knee joint) is used. Reproduce the load stimulus
Thereby, while being able to reproduce easily the force concerning the cell in the state enclosed by the matrix which is an extracellular tissue, the culture body C can be cultured in the state approximated to the environment of the structure | tissue in the biological body. Therefore, the adaptability to the living body after transplantation of the culture can be improved.
In this case, the culture body C composed of a structure containing cultured tissue or cultured cells can be formed in advance in a shape suitable for transplantation. Specifically, a matrix of the structure containing cultured cells is used as a biocompatible material. For example, a cell support mainly composed of collagen can be used, and a structure including cultured cells in which chondrocytes or meniscal cells are seeded and adhered can be used.

  In addition, the other structure and effect | action of this biomechanical stimulation load apparatus are the same as that of the biomechanical stimulation load apparatus of the said 1st Example and 2nd Example.

The present invention sterilizes the inside of a carbon dioxide gas incubator and cultures composed of cultured cells, cultured tissues or structures containing cultured cells in an in vitro culture in an environment in which temperature, humidity, oxygen, carbon dioxide, nitrogen partial pressure, etc. are controlled. The body is subjected to a biomechanical stimulus load in an in vivo environment, an environment similar to the structure, a vertical load load stimulus using gravity, and a lateral shear stress stimulus in the horizontal plane It can be applied directly by vibrating.
In addition, the biomechanical stimulation load device of the present invention can be operated from the outside of the carbon dioxide incubator 2, and the culture medium can be replaced by an external operation, so that continuous culture can be performed for a long time. . Further, this biomechanical stimulation load device does not require a large amount of power, and it is possible to manufacture a biomechanical stimulation load device that is accurate and compact in load weight.
As a result, the culture can be cultured in a state that approximates the tissue environment in the living body, so that it becomes possible to create a differentiated and mature culture that can withstand the mechanical load after transplantation. Can be performed efficiently.
In addition to performing in vitro biomechanical stress stimulation that was not possible with conventional tissue culture techniques, culturing the culture in a form suitable for transplantation has an effect on the growth and differentiation of the culture and is suitable for transplantation. An organization can be created.
In addition, by the basic medical research using the biomechanical stimulus loading device of the present invention , tissue that has been subjected to biomechanical load stimulation outside the body, it is possible to create a differentiated and matured cultured tissue that can withstand the mechanical load after transplantation, It can contribute to tissue regeneration and transplantation medicine.
Furthermore, since the present invention can apply stimuli very similar to the standing, walking, running, jumping, and twisting of the living body, it can contribute to the elucidation of the mechanism of destruction and damage of bone, cartilage, skin, etc. I can expect. Specifically, the weight of the load piston in the vertical direction can be changed to the weight of the damaged load, the descending speed can be increased and a shocking stimulus in the vertical direction can be applied. It is also possible to apply a stimulus in a state where the knee is twisted by applying a shear stress of. It is also possible to study bone wear, etc. by repeating lateral movement and applying shear stress for a long time.

  The biomechanical stimulation load device of the present invention can be subjected to mechanical stimulation load under various conditions as a tissue culture device, and can be miniaturized with a relatively simple device. There is an unprecedented advantage in that a stress similar to the environment in a living body can be applied by applying a shear stress stimulus via an extracellular matrix. It can be widely used as a product of standard culture equipment in laboratories in the field of tissue regeneration in medical biology, and there is a possibility of new industrialization. Or if it is recognized that it is a useful device for culturing structures containing cultured cells, and if it is a production device for cartilage, the need for regenerative treatment of the meniscus alone is equal to or better than that of bone In Japan alone, 10,000 cases can be the target year.

BRIEF DESCRIPTION OF THE DRAWINGS It is whole explanatory drawing which shows 1st Example of the biomechanical stimulation load apparatus of this invention. It is the principal part explanatory drawing. It is explanatory drawing which decomposed | disassembled which shows the piston for load loads. It is explanatory drawing which shows an internal culture container and a receiving group. It is whole explanatory drawing which shows 2nd Example of the biomechanical stimulation load apparatus of this invention. It is the principal part explanatory drawing. It is explanatory drawing of the osteochondral structure of the knee joint of the meniscus tissue culture example which shows 3rd Example of the biomechanical stimulus loading apparatus of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Carbon dioxide incubator 2 Mechanical stimulus load device 20 Guide member 21 Piston vertical movement stage 21a Hole part 22 Stage raising / lowering mechanism 22a Electric actuator 22b Wire 23 Piston mounting stage 23a Hole part 23b Piston mounting stage drive mechanism 24 Culture Container fixing stage 25 Shear stress stage 26 Shear stress loading mechanism 27 Additional load heavy restraint stage 28 Additional load heavy restraint stage 28b Additional load heavy restraint stage drive mechanism 3 Culture container 30 Internal culture container 31 Receptor 31A Artificial Tibial bone cartilage 32 Medium tank (for injection)
33 Medium tank (for discharge)
4 Control computer 5 Piston for load application 5a Piston for load application 5b Piston for load application 51 Pressurization body 51A Pressurization body 51Aa Pressurization body 51Ab Pressurization body 51C Pressurization body 51D Artificial femur bone cartilage 52 Guide shaft part 53 Large-diameter portion 54 Heavy load attachment portion 55 Additional load heavy load 55a Additional load heavy load 55b Additional load heavy load C Culture body (cultured cell, cultured tissue or structure containing cultured cell)
X Cultured body load load stimulation part

Claims (9)

  A biomechanical stimulus load device comprising a mechanical stimulus load device and a culture vessel installed in a carbon dioxide incubator, and a control computer, the mechanical stimulus load device comprising a load load piston and the load load A piston vertical movement stage that supports the piston for movement in the vertical direction within a predetermined range, and a stage lifting mechanism for moving the piston vertical movement stage in the vertical direction. By driving the mechanism in a predetermined cycle, the piston vertical movement stage is moved in the vertical direction so that the weight of the load loading piston is applied to the culture body in the culture vessel when the piston vertical movement stage is lowered. A biomechanical stimulation load device, wherein the piston mounting stage is mounted on the piston vertical movement stage. A load loading piston is inserted into and supported by a plurality of holes formed in the piston mounting stage, and a plurality of loads are applied to the piston vertical movement stage. A biomechanical stimulus loading device, characterized in that a common hole for allowing the loading piston to freely move in the vertical direction is formed.   2. The biomechanical stimulus loading device according to claim 1, wherein an additional load weight is attached to an upper end portion of the load load piston.   3. The biomechanical stimulus loading device according to claim 1, wherein a pressurizing body having a shape corresponding to the culture body is attached to a lower end portion of the load loading piston. Claim piston mounting stage adapted to support a plurality of load-bearing piston, the piston vertical moving stage, characterized in that so as to movable in a horizontal plane 1,2 Or the biomechanical stimulus loading device according to 3.   5. The load loading piston is supported by a piston vertical movement stage or a piston mounting stage so that the piston does not rotate when moving in the vertical direction. Biomechanical stimulus loading device.   6. The biomechanical stimulus loading device according to claim 1, wherein the culture vessel is detachably attached to a culture vessel fixing stage of the mechanical stimulus loading device.   7. The biomechanical stimulus loading device according to claim 1, further comprising a shear stress loading mechanism for moving or vibrating the culture vessel in a horizontal plane.   A culture-body-load-stimulation unit consisting of a pressurizing body and a reception group at the lower end of the load-loading piston can be fixed in the culture vessel. 8. The biomechanical stimulation load device according to claim 1, wherein the biomechanical stimulation load device has a similar structure.   A member made of a biocompatible synthetic resin material is attached to a culture body load-stimulating portion composed of a pressurizing body and a receiving group at a lower end portion of a load-loading piston. 9. The biomechanical stimulus loading device according to 8.

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